bluuebunny/crossref_metadata_2025 huggingface dataset:
FORMATION OF A HEMOLYTICALLY ACTIVE CELLULAR INTERMEDIATE BY THE INTERACTION BETWEEN PROPERDIN FACTORS B AND D AND THE ACTIVATED THIRD COMPONENT OF COMPLEMENT
Authors: Fearon Douglas T. , Austen K. Frank , Ruddy Shaun
Published: June 2004

The present studies demonstrate that the factor B-dependent C3 convertase can be affixed to an erythrocyte by use of an intermediate bearing C3b and that this convertase brings the hemolytic reaction to completion with an efficiency comparable to that of classical convertase. The evidence that the EAC43 intermediate was lysed by a new pathway includes requirements for factors B and D and cell-bound C3b for subsequent lysis by the terminal components, C3-C9. The linear stoichiometry of the effective molecule titrations of C3b and factor B, and the first-order kinetics displayed by the generation and decay of the factor B-dependent hemolytic site are characteristics consistent with the one-hit theory as initially developed for the classical complement system. The use of hemolytically active cellular intermediates to examine the reactions occurring with C3b and factors B and D has allowed extension of the one-hit theory to this molecular sequence, development of effective molecule titrations, recognition of the analogies to the functional characteristics of the classical C3 convertase, and discrimination of the probable mechanism of terminal complement activation from reactive lysis.
ADAPTATIONS OF ENERGY METABOLISM IN THE CULTIVATED MACROPHAGE
Authors: Simon Lawrence M. , Axline Stanton G. , Horn Barry R. , Robin Eugene D.
Published: June 2004

Adaptive changes in energy metabolism, as reflected by pyruvate kinase and cytochrome oxidase activities, were examined during in vitro differentiation of the cultivated macrophage. Serum concentrations of tissue culture media, which directly influence endocytic activity, and ambient oxygen tension were both shown to influence pyruvate kinase and cytochrome oxidase activities. Cells maintained in high serum concentrations (30% newborn calf serum [NBCS]) exhibited a 300–400% increase in pyruvate kinase activity and a 40% increase in cytochrome oxidase activity, whereas cells maintained in low serum concentrations (2% NBCS) exhibited a lesser increase (65%) in pyruvate kinase activity and no change in cytochrome oxidase activity. Anaerobiosis resulted in additional alterations in pyruvate kinase and cytochrome oxidase activities. Cells maintained for 48–72 h under anaerobic conditions exhibited a 500–600% increase in pyruvate kinase activity and a 40% decrease in cytochrome oxidase activity. Increased pyruvate kinase activity was dependent on continued protein synthesis. Enzyme increases occurred in anaerobically cultured cells despite an overall reduction in cell protein synthesis. It is suggested that adaptive changes in pyruvate kinase and cytochrome oxidase activity resulting from alterations in either serum concentration or ambient oxygen tension are regulated by two independent mechanisms. One mechanism is aimed at providing energy for endocytic activity and the other in compensating for impaired oxidative metabolism during anaerobiosis.
ANATOMICAL DISTRIBUTION OF T AND B LYMPHOCYTES IN THE RAT
Authors: Goldschneider Irving , McGregor D. D.
Published: June 2004

A method is described whereby antisera raised in rabbits to rat thoracic duct lymphocytes were made specific for the plasma membrane antigens of T and B lymphocytes. These lymphocyte-specific antisera were used in immunofluorescence assays to study the distribution of B and T cells in lymphocyte containing tissues and body fluids of the rat.

Rabbit antirat B-cell serum (ALSB) reacted selectively with the surfaces of lymphocytes in the lymphoid follicles of lymph node cortex and in the follicles and marginal zones of splenic white pulp, but not with the surfaces of germinal center cells or plasma cells. An identical pattern of fluorescent staining was obtained with rabbit antirat Ig serum. It was shown by blocking, absorption, and immunoprecipitation studies that ALSB was composed in large part of antibodies to rat Ig, but that it contained antibodies to other B-cell antigens as well.

Rabbit antirat T-cell serum (ALST) reacted selectively with the surfaces of lymphocytes in the paracortex of lymph node and in the periarteriolar sheath of spleen, and with thymocytes. ALST did not display anti-Ig activity.

ALST reacted with approximately 100% thymocytes and with 90% thoracic duct, 80% lymph node, 60% blood, 50% spleen, and 10% bone marrow lymphocytes in suspensions of cells from these sources. ALSB reacted with the remainder of the lymphocytes in the suspensions, except for bone marrow in which only 59% of lymphocytes had detectable B- or T-cell surface antigens.

The population of T lymphocytes in rat bone marrow was depleted by drainage of lymphocytes from a thoracic duct fistula, thereby establishing their membership in the pool of recirculating T cells. Approximately 14% of lymphocytes issuing from the thoracic duct of TxBM donors reacted with ALST. The presence in these animals of a small number of T cells, calculated to be approximately 2% of the normal value, may account for the limited capacity of TxBM rats to respond to antigens that induce a cell-mediated immune response.
EFFECT OF RECENT ANTIGEN PRIMING ON ADOPTIVE IMMUNE RESPONSES
Authors: Sprent J. , Miller J. F. A. P.
Published: June 2004

When thoracic duct lymphocytes (TDL) or mesenteric lymph node (MLN) cells from mice primed 1 day before with either sheep erythrocytes (SRC) or horse erythrocytes (HRC) were transferred together with both SRC and HRC to irradiated mice, antibody responses measured 7 days later could not be detected to the priming antigen but were high to the other antigen. Furthermore, this unresponsiveness of TDL and MLN to the priming antigen could not be abrogated by delaying antigen challenge of the transferred cells for 1–2 wk.

Previous work had shown that short-term priming with antigen also induced specific unresponsiveness in spleen cells on adoptive transfer. Unresponsiveness in these cells, however, was only of temporary duration, full recovery in the reactivity of the cells being observed when challenge with the priming antigen on transfer was delayed for 5 or more days. Since the present work showed that such recovery from initial unresponsiveness on transfer was unique to spleen cells and did not apply to TDL or MLN, it appeared that different mechanisms were responsible for the unresponsiveness in the three populations. It is proposed that the unresponsiveness detected in TDL and MLN cells in the present study resulted from a deficiency of antigen-reactive cells, these cells having been recruited to the spleen, i.e., a region of antigen concentration. This concept of antigen-induced selective recruitment of circulating lymphocytes was supported by evidence that 51Cr-labeled heterologous erythrocytes indeed localized largely in the spleen after intravenous injection but not in MLN.
THE THYMIC SUPPRESSOR CELL
Authors: Ha Tai-You , Waksman Byron H. , Treffers H. Peter
Published: June 2004

This study confirms the finding that washed thymocytes from rats given 100 mg of BGG 48 h earlier, when transferred to syngeneic recipients, exert a specific suppressor effect on immunological responses to BGG in the latter. The active cells are found in a subpopulation of low density, making up less than 10% of the total thymocytes, and are partially resistant to hydrocortisone.
CLONAL NATURE OF THE IMMUNE RESPONSE TO PHOSPHORYLCHOLINE
Authors: Claflin J. Latham , Lieberman Rose , Davie Joseph M.
Published: June 2004

The relationship between receptor molecules on antigen-binding lymphocytes (ABC) and antibody produced by antibody-secreting cells was studied in inbred strains of mice using the immune response to phosphorylcholine (PC) as a model system. Splenic and lymph node lymphocytes of nonimmune mice possess rare lymphocytes which bind 125I-labeled PC-bovine serum albumin. The frequency of PC-ABC increases after immunization and is paralleled by a rise in the frequency of PC-specific antibody-producing cells. Both of these responses are thymus independent. The receptors on these ABC display specificity for PC and are exclusively of the IgM class. In one of the strains, BALB/c, the receptors possess the same idiotype and fine degree of specificity for PC and two of its analogues, glycerophosphorylcholine and choline, that are characteristic of a PC-binding myeloma, HOPC 8. Furthermore, the idiotype and class of the receptor in these mice do not change during the course of the immune response. These data provide more direct evidence for the immunelogic relevance of receptor-bearing lymphocytes.
CYTOTOXIC IMMUNE CELLS WITH SPECIFICITY FOR DEFINED SOLUBLE ANTIGENS
Authors: Schirrmacher Volker , Rubin Bent , Pross Hugh , Wigzell Hans
Published: June 2004

Spleen cells from mice immunized against ovalbumin (OA) or dinitrophenylated mouse serum albumin (DM) were found to be specifically cytotoxic in vitro towards target cells (chicken red blood cells) coated with these antigens. Inhibition of specific cytotoxicity was observed when free soluble antigen was added to the incubation mixtures. DM-immune cell cytotoxicity could be specifically and completely inhibited by DNP-lysine and was thus shown to be hapten specific. Complete and specific inhibition was also observed for OA-immune cell cytotoxicity using OA as inhibitor, but compared with the inhibition curves obtained with DNP-lysine, the OA cytotoxicity inhibition curves were shifted by a factor of about one hundred towards lower molar inhibitor concentrations. Very similar results were observed when the serum antibodies of DM- and OA-immune animals were analyzed by passive hemagglutination inhibition. With increasing time after immunization, both cytotoxicity inhibition curves and agglutination inhibition curves, shifted to lower antigen or hapten concentrations.

Specific cytotoxicity against antigen-coated target cells was induced in nonimmune spleen cells (a) by serum from immune animals, and (b) by supernatants from in vitro immune cell cultures. In both instances, the factor which induced antigen-specific cytotoxic activity could be absorbed on anti-mouse Ig columns, thus demonstrating its immunoglobulin nature. The ability of target cell bound antibodies to induce cytotoxicity in nonimmune spleen cells was restricted to the 7S antibody class.
SEGMENTAL RESPONSE OF THE MACROPHAGE PLASMA MEMBRANE TO A PHAGOCYTIC STIMULUS
Authors: Griffin Frank M. , Silverstein Samuel C.
Published: June 2004

A method of attaching mouse RBCs to mouse macrophages is described. Both cell types were coated with rabbit anti-mouse macrophage F(ab')2, and cross-linkage of cells was effected with sheep F(ab')2 directed against rabbit F(ab')2. 98% of macrophages attached an average of 11 RBCs each. Attachment occurred at 37°C and was stable for at least 4 h. Less than 0.1% of macrophages ingested RBCs under these conditions.

Latex particles and opsonized pneumococci were ingested as avidly by RBC-coated macrophages as by native macrophages. Ingestion of these particles did not prompt ingestion of attached RBCs. When anti-RBC IgG was added, however, over 90% of macrophages ingested an average of six RBCs each. Thus, ingestion of one particle does not trigger generalized phagocytosis of all particles attached to the cell's plasma membrane, and the phagocytic stimulus is confined to the segment of the cell's plasma membrane immediately adjacent to the particle being ingested.
FUNCTIONAL CHARACTERISTICS OF PEYER'S PATCH LYMPHOID CELLS
Authors: Kagnoff Martin F. , Campbell Stephen
Published: June 2004

Peyer's patches from normal mice contain antigen-sensitive B and T cells, but lack the accessory adherent cell type(s) required both for the induction of humoral immune responses and for the induction of allograft reactions against cell surface alloantigens. Immune responsiveness can be restored to cultures of Peyer's patch cells by the addition of either APEC or ME. Peyer's patch B cells can be specifically induced by antigen to synthesize humoral antibody. Peyer's patch T cells can cooperate in B-cell induction and can be induced to mediate an allograft reaction against an allogeneic stimulus. Peyer's patch lymphoid aggregates appear to be a storehouse of antigen-sensitive cells sequestered in such a way as to lack an accessory cell type or factor required for induction,
GENETIC INFLUENCES AFFECTING THE OCCURRENCE OF A DIABETES MELLITUS-LIKE DISEASE IN MICE INFECTED WITH THE ENCEPHALOMYOCARDITIS VIRUS
Authors: Craighead John E. , Higgins Duncan A.
Published: June 2004

The M variant of encephalomyocarditis virus produces a diabetes mellitus-like disease in DBA/2 mice but not in animals of the C3H strain. Fewer than one-third of infected F1 (DBA/2 x C3H) progeny exhibit the disease, whereas the prevalence in backcrosses (F1 x DBA/2, F1 x C3H) is comparable to the parental inbred strain. Thus, the mode of inheritance of the diabetic predisposition appears to be polygenic. DBA/2 animals develop striking inflammatory and necrotizing lesions of the islets of Langerhans; in contrast, alterations of the insular tissue in the C3H mice are minimal. Although metabolic abnormalities appear to be consequent to lesions of beta cells, the factors influencing the severity of these insular changes are incompletely understood.
INFLUENCE OF DOSE AND ROUTE OF ANTIGEN INJECTION ON THE IMMUNOLOGICAL INDUCTION OF T CELLS
Authors: Lagrange P. H. , Mackaness G. B. , Miller T. E.
Published: June 2004

Delayed-type hypersensitivity (DTH) develops in the absence of an adjuvant when mice are injected intravenously or subcutaneously with an appropriate dose of sheep red blood cells (SRBC). The optimal intravenous dose of 105 SRBC (in CD-1 mice) produces maximum DTH which decays exponentially from its peak on day 4. Increasing the dose of SRBC reduces and eventually abolishes all evidence of DTH. DTH fails to reappear in respose to secondary stimulation except in splenectomized mice in whom the development of DTH is not suppressed, even by massive doses of SRBC. Hence the suppression cannot be due to antigen as such.

The optimal dose of SRBC for sensitization by footpad inoculation is 100-fold higher (107 SRBC in CD-1 mice), but even 109 SRBC do not block the induction of DTH by this route of immunization. A blocking dose of SRBC, given intravenously 1 day before footpad inoculation, completely suppresses cell proliferation in the draining lymph node, prevents PFC production there, and blocks the induction of DTH by a sensitizing dose of SRBC. If given 1 day after footpad sensitization, intravenous antigen has little effect on the cellular response in the regional node but DTH is still completely suppressed. Blocking of induction and expression may depend, therefore, on different mechanisms.
RESTRICTION OF GENE EXPRESSION IN B LYMPHOCYTES AND THEIR PROGENY
Authors: Jones Patricia P. , Cebra John J. , Herzenberg Leonard A.
Published: June 2004

Lymphocytes from b5/b9 rabbits were stained in suspension with fluorescent antiallotype antibody reagents to selectively label with fluorescent molecules those cells bearing membrane immunoglobulin (Ig) of the b5 or b9 allotype. After staining, the cells were separated by the fluorescence-activated cell sorter into populations markedly enriched in cells bearing b5 or b9 membrane Ig or totally depleted of cells with detectable membrane Ig. The potential of these separated cells to give rise to Ig-synthesizing plasma cells either in vivo after transfer into irradiated recipients or in vitro during culture in the presence of phytohemagglutinin or pokeweed mitogen was assessed by immunofluorescence. The relative proportion of b5 and b9 cytoplasmic Ig-stained cells (CSC) arising from the separated cells was determined to test directly whether B lymphocytes and their progeny are committed to the synthesis of Ig of one allotype.

It was found that b5- and b9-bearing cells gave rise almost exclusively to b5- and b9-producing plasma cells, respectively, in both the in vivo and in vitro assay systems. Most of these CSC were probably not derived from previously existing CSC but arose as the result of the differentiation of lymphocytes with membrane Ig. When cell populations totally depleted of Ig-bearing lymphocytes were cultured, very few CSC were found, indicating that the majority of immediate precursors of CSC have membrane Ig. These results suggest that individual B cell clones are phenotypically restricted to the expression of immunoglobulin genes on one chromosome; the significance of this clonal allelic exclusion is discussed.
THE INTERACTION OF IgE WITH RAT BASOPHILIC LEUKEMIA CELLS
Authors: Kulczycki Anthony , Isersky Chaviva , Metzger Henry
Published: June 2004

A rat basophilic leukemia cell line originally described by Eccleston et al. (3) has been successfully transplanted into eight Wistar strains and adapted to suspension cell culture without noticeable morphological changes.

The cells deplete the reaginic activity of mouse and rat immune sera to an extent roughly equivalent to that reported for normal rat mast cells. Studies utilizing radioiodinated antilight-chain antibodies and radioiodinated partially purified rat IgE indicate that the cells bind IgE to their surface membrane with high specificity. Heating or mildly reducing the rat IgE destroyed its binding activity. The binding is unaffected by the presence or absence of Ca++ and Mg++, and is markedly inhibited by reaginic but not by normal rat or mouse serums. The affinity of these cells for human IgE, if present at all, is substantially lower than for rat IgE.
RECEPTOR FOR SOLUBLE C3 AND C3b ON HUMAN LYMPHOBLASTOID (RAJI) CELLS
Authors: Theofilopoulos Argyrios N. , Bokisch Viktor A. , Dixon Frank J.
Published: June 2004

This study describes the presence of a receptor for fluid phase human C3 and C3b on Raji cell membranes. The binding of C3 and C3b was demonstrated indirectly by a fluoresceinated anti-C3 serum and directly by using radioiodinated proteins. No other complement proteins or serum factors were needed to mediate binding of C3 and C3b to the receptor. The possibility of enzymatic cleavage of C3 before or after its attachment on the cell membrane was ruled out by the demonstration of antigenically intact C3 on Raji cells. Inhibition and dissociation of Raji cell-EAC1423 rosettes by C3 and C3b indicated that both of these proteins bind to the same receptor site or closely associated receptor sites on Raji cells. C3b-bearing Raji cells were immune adherence negative, indicating that C3b binding to the receptor is brought about through the immune adherence region of the molecule and not the C3d portion. The C3 receptor on Raji cell membranes is uniformly distributed and can move on the membrane plane. Approximately 4 x 105 molecules of C3 or C3b bind per Raji cell. The receptor had a higher affinity for C3 than C3b, as was shown by uptake experiments and inhibition of Raji cell-EAC1423 rosette formation. Apart from the described receptor for C3 and C3b another specific receptor for C3b inactivator-cleaved C3b (C3d) bound to red cells was shown to be present on Raji cells.

Raji cells cultured in medium containing fresh normal human serum and cobra venom factor were lysed. Similar results were obtained when C3b-bearing Raji cells were cultured in medium with fresh normal human serum. The lytic effect could be abolished by inactivating serum C3 proactivator (C3PA) and required C6. It was concluded that C3b bound to the Raji cell membrane activates the complement system through the alternate pathway and results in membrane damage and cytolysis. It is postulated that cell destruction by this mechanism may play an important role in vivo in controlling cell growth.
NEW, THIRD CLASS OF AMYLOID FIBRIL PROTEIN
Authors: Husby G. , Natvig J. B. , Sletten K.
Published: June 2004

An unusual protein AR was isolated from the amyloid fibril preparation derived from a patient with primary amyloidosis. Protein AR was unique in its antigenicity, and revealed no structural identity with any known amyloid proteins or with immunoglobulin chains or fragments. Thus a new third class of amyloid fibril proteins besides the immunoglobulin light-chain variable region fragments and the nonimmunoglobulin protein AS, has been characterized. A component antigenically related to protein AR was found in the serum of the patient.
THE EFFECT OF SEA STAR COELOMOCYTE EXTRACT ON CELL-MEDIATED RESISTANCE TO LISTERIA MONOCYTOGENES IN MICE
Authors: Willenborg David O. , Prendergast Robert A.
Published: June 2004

Mice treated with sea star factor (SSF), a protein extracted from sea star coelomocytes, became highly susceptible to infection with a normally sublethal dose of Listeria monocytogenes. This was in contrast to the expected result of increased resistance originally postulated because of the macrophage-activating properties of SSF. Enhanced susceptibility was seen when SSF was given from 96 h before to 48 h after infection with Listeria. Mice pretreated with SSF failed to develop immunity to Listeria when given a dose of organisms capable of immunizing nontreated mice. Treatment of immune mice with SSF did not alter their immune status. In addition, incubation of immune lymphocytes with SSF in vitro did not alter their ability to adoptively transfer immunity to normal recipients. Immune lymphocytes treated with SSF and then incubated with anti-SSF and C did, however, lose the ability to transfer immunity. These results suggest that SSF enhances infection by binding to T lymphocytes, inhibiting their replication upon contact with Listeria antigen and thus preventing the generation of a population of sensitized lymphocytes capable of effecting anti-Listeria immunity.
MIXED LYMPHOCYTE CULTURE RESPONSES OF MICE
Authors: Plate Janet M. D.
Published: June 2004

Cellular responses in vitro to H-2D region histocompatibility antigens were demonstrated to be under the genetic control of two or three (P = 0.013) independently segregating loci. The H-2 region itself accounts for one of these loci, however, its activity appears to be dependent upon an association with other non-H-2-associated genetic information. The ability to stimulate a response and to respond to that stimulus are two separate genetic functions in certain MLR combinations. The stimuli in our studies were products of the H-2D region and cell donors must differ at that region in order for a response to occur. The control of the level of responses was determined by other genetic material. Differences at these "response loci" were not necessary for the induction of a proliferative response in the mixed lymphocyte cultures.
CELL-BOUND RECEPTORS FOR ALLOANTIGENS ON NORMAL LYMPHOCYTES
Authors: Binz Hans , Lindenmann Jean , Wigzell Hans
Published: June 2004

Antialloantibodies were prepared in F1 hybrid rats by immunization with alloantibodies from one parent raised against antigens of the other parent. The Ig fraction of such antialloantibodies was iodinated and used to investigate the nature of idiotype-carrying normal lymphoid cells. Lymphoid cell populations of the proper genotype fixed radioactive antialloantibody in proportion to their B-cell content. Activated T-cell populations, when depleted of B cells, did not fix significant amounts of radioactivity. Idiotype-carrying cells were sensitive to rabbit antirat Ig serum lysis and to antialloantiserum lysis, but not to rabbit anti-T-serum lysis. Also, the normal idiotype-containing B cell could be shown to have the expected antigen-binding specificity of its receptor. This was shown by the use of fibroblast cell monolayers that adsorbed specifically those cells capable of fixing the proper antialloantibody.
INHIBITION OF SPECIFIC CELL-MEDIATED CYTOTOXICITY BY ANTI-T-CELL RECEPTOR ANTIBODY
Authors: Kimura Arthur K.
Published: June 2004

The present study describes a method for the production of a specific anti-T-cell receptor antiserum, and characteristics of its ability to block specific cell-mediated cytotoxicity in vitro. Immunization and antiserum adsorption procedures were designed to select for idiotypic differences in the recognition units of C3H lymphocytes immune to two different strains of mouse cells, such that the reactivity of only one population of effector cells is inhibited by this antiserum. Both in vivo and in vitro sensitized effector T cells are subject to this inhibition. That the site of the antiserum blockade is clearly on the effector cell and not on the target cell is demonstrated.
DEMONSTRATION OF A BLOCKING FACTOR IN THE PLASMA AND SPINAL FLUID OF PATIENTS WITH SUBACUTE SCLEROSING PANENCEPHALITIS
Authors: Ahmed Aftab , Strong Douglas M. , Sell Kenneth W. , Thurman Gary B. , Knudsen Richard C. , Wistar Richard , Grace William R.
Published: June 2004

Conflicting reports on the immune responsiveness of patients with subacute sclerosing panencephalitis (SSPE) have been reported. This report shows that the leucocytes from four SSPE patients exhibited strong sensitivity to both measles and SSPE virus preparations as measured by the macrophage migration inhibition test, mixed lymphocyte virus infected cell culture test, and the lymphotoxin assay. Earlier suggestions that a factor may be operating to suppress cellular reactivity are confirmed by the demonstration that the response of lymphocytes from SSPE patients could be blocked by the addition of SSPE spinal fluid or plasma. It was determined that the blocking factor was stable at –20°C, heat labile at 56°C for 30 minutes, trypsin and neuraminadase sensitive, and had a mol wt greater than 150,000 as determined by Sephadex G-200 gel chromatography. The blocking factor appeared to be specific for SSPE virus and did not block the response of lymphocytes to nonspecific mitogenic agents and other viral and bacterial agents.
COMMON CELL SURFACE ANTIGEN ASSOCIATED WITH MAMMALIAN C-TYPE RNA VIRUSES
Authors: Yoshiki Takashi , Mellors Robert C. , Hardy William D. , Fleissner Erwin
Published: June 2004

The indirect membrane immunofluorescence test and the absorption analysis of rabbit anti-FeLV, rabbit anti-FeLVp 30, and rabbit anti-MuLVp 30 antisera yielded the following conclusions. An antigen shared by mammalian (murine and feline) C-type RNA leukemia and sarcoma viruses was detected on the surface of cells infected or transformed by C-type viruses. The antigen was characterized as membrane-bound gs antigen bearing two determinants, membrane-bound gs-1, intraspecies-specific antigenic determinant, and membrane-bound gs-3, interspecies-specific antigenic determinant. Membrane-bound gs antigen was located on the cell surface, frequently near the site of virus budding but not on the envelope of murine C-type RNA virus.
STIMULATION OF ANTIBODY PRODUCTION TO THE HAPTEN 2,4-DINITROBENZENE BY AFFINITY LABELED MURINE LYMPHOID CELLS
Authors: Lawrence David A. , Weigle William O.
Published: June 2004

The ability of meta-nitrobenzenediazonium fluoborate (m-NBDF)-labeled thymus and spleen (S) cells to transfer immunity to 2,4-dinitrophenyl (DNP) into irradiated syngeneic recipients was investigated. There was a significant increase in the number of anti-DNP plaque-forming cells (PFC) when m-NBDF-labeled thymus cells and normal spleen cells, or normal thymus cells and m-NBDF-labeled spleen cells were transferred, but not when both thymus- and S-cell populations were labeled and injected together into irradiated recipients. The ability of these cell populations to cooperate and enhance the in vivo immune response to DNP is discussed. The T cells seem to be actively involved in the development of this response; they participate beyond the mere role of carrying and presenting antigen to the B cells. It is suggested that cell to cell contact between T and B cells may be an important factor in the elicitation of an immune response. In addition, the cellular interaction is affected by irradiating the thymus cell preparation and the initiating interaction required for antibody synthesis probably occurs within 48 h after injecting the cell populations into the syngeneic irradiated recipients.
THE ROUTE OF ENTERIC INFECTION IN NORMAL MICE
Authors: Carter Philip B. , Collins Frank M.
Published: June 2004

This study followed the early pathogenesis of orally induced murine typhoid fever. Intragastrically administered Salmonella enteritidis moves quickly through the normal undisturbed gut so that only a small residuum remains in the cecum and large intestine after the first few hours. Dye injection of the gut wall was used to show that lymph from discrete portions of the gastrointestinal tract drains to separate lymph nodes, probably via the regional Peyer's patches. Plating techniques capable of detecting a single colony-forming unit of S. enteritidis within the different Peyer's patches and draining lymph nodes indicate that, although the cecum and large intestine are exposed to large numbers of Salmonella for longer time periods than the small intestine, the primary site of bacterial penetration involves the distal ileum. This area of the small intestine as well as the cecum are both drained by the distal mesenteric lymph nodes, and were the only nodes which contained detectable numbers of viable Salmonella over the first 24 h of infection. Neither the pyloric nor the proximal mesenteric lymph nodes (which drain the stomach and duodenum) nor the pancreatic and caudal lymph nodes (which drain the transverse and descending colon) contained viable Salmonella.

Salmonella were observed to infect the ileal mucosa and its Peyer's patches. With time, this infection progresses to the draining lymph node and ultimately reaches the liver and spleen. Some of the implications of these findings relative to the development of acquired resistance to enteric disease are discussed.
A SERUM FACTOR IN CHRONIC HYPOCOMPLEMENTEMIC NEPHRITIS DISTINCT FROM IMMUNOGLOBULINS AND ACTIVATING THE ALTERNATE PATHWAY OF COMPLEMENT
Authors: Vallota Enrique H. , Götze Otto , Spiegelberg Hans L. , Forristal Judith , West Clark D. , Müller-Eberhard Hans J.
Published: June 2004

Nephritic factor (C3NeF) has been isolated from plasma of patients with hypocomplementemic chronic glomerulonephritis (HCG) by ion exchange and molecular sieve chromatography. This material was further treated with solidified anti-Ig antiserum. The purified material failed to react with antiserum to human IgG, IgG3, Fab, Fc, and kappa and lambda chains, but retained full C3NeF activity. The nonidentity of C3NeF with IgG was further demonstrated by Ouchterlony analysis using anti-IgG and anti-C3NeF. Isolated C3NeF was found to be a protein with a sedimentation coefficient of 7S and a mol wt of 150,000 daltons, which on microzone electrophoresis and gel electrophoresis at pH 8.6 behaved as a γ-globulin. C3NeF is not a C1q precipitin and does not activate the classical complement pathway. Unlike cobra venom factor, it failed to enter into a complex with C3 proactivator (C3PA) when incubated with normal human serum (NHS) and then subjected to sucrose density gradient ultracentrifugation. The action of isolated C3NeF on C3 requires C3PA, C3PA convertase (C3PAse), and properdin (P). Similarly, C3PA conversion by C3NeF requires P, C3PAse, and C3. Total hemolytic activity was lost by incubation of 64 µg of C3NeF/1 ml NHS at 37°C for 30 min. Both C3a and C5a anaphylatoxin could be generated by C3NeF in serum previously depleted of anaphylatoxin inactivator. Anti-C3NeF was found to detect an antigen in all NHS tested. Treatment of NHS with solidified anti-C3NeF caused impairment of the alternate complement pathway. It failed to sustain lysis of glutathione-treated human erythrocytes initiated by inulin. It is conceivable that the normal serum constituent which is removed by anti-C3NeF constitutes the inactive precursor of C3NeF, and a heretofore unrecognized component of the alternate pathway.
ALTERED FUNCTIONAL PROPERTIES OF THE RENAL GLOMERULUS IN AUTOLOGOUS IMMUNE COMPLEX NEPHRITIS
Authors: Schneeberger E. E. , Leber P. D. , Karnovsky M. J. , McCluskey R. T.
Published: June 2004

The altered functional properties of the glomerular capillary wall in a model of autologous immune complex disease (Heymann's nephritis) was studied by electron microscopy using intravenously injected protein tracers of varying molecular weight. There was an increase in the permeability of the glomerular basement membrane (GBM) itself to large molecules; this change was focal and was found in those areas where the GBM contained immune complex deposits. Both ferritin and catalase, tracers normally restricted from passing the glomerular filter, were present in the urinary space within minutes of injection. No evidence was obtained for increased glomerular epithelial transport in this disease. Foot process swelling and "close" junction formation was moderate, even in animals with marked degrees of proteinuria. Indirect evidence, therefore, makes an alteration in the slit pore complex likely. In addition, there was immediate and selective concentration of all tracers within deposits, though ferritin was partially excluded from some deposits. This phenomenon may be of significance in the perpetuation of the disease.
HORMONAL CONTROL OF LYSOSOMAL ENZYME RELEASE FROM HUMAN NEUTROPHILS
Authors: Ignarro L. J. , Lint T. F. , George W. J.
Published: June 2004

The purpose of this investigation was to examine the effects of autonomic neurohormones, cyclic nucleotides, and related agents on the immunologic discharge of lysosomal enzymes from, and phagocytosis by, purified human neutrophils. In order to discern the possible intracellular mechanisms by which certain neurohormones influence neutrophil function, the concentrations of cyclic AMP and cyclic GMP in neutrophils were assessed during cell contact with phagocytizable particles and autonomic agents. The model system employed for study was the interaction of purified human neutrophils with rheumatoid arthritic (RA) serum-treated zymosan particles at 37°C in a neutral, balanced salt solution containing glucose. Neutrophils ingested the particles and discharged ß-glucuronidase but not lactate dehydrogenase activity during 30 min of incubation. Treatment of zymosan particles with RA serum was more effective than treatment with normal serum with regard to the extent of both particle uptake and lysosomal enzyme release. During contact of neutrophils with RA serum-treated zymosan particles epinephrine, isoproterenol, and cyclic AMP inhibited both particle ingestion and ß-glucuronidase discharge. These actions of epinephrine were associated with a concomitant elevation of cyclic AMP levels. In contrast to the actions of catecholamines and cyclic AMP, acetylcholine and cyclic GMP accelerated lysosomal enzyme release without affecting particle uptake. The actions of acetylcholine were associated with a concomitant elevation of cyclic GMP levels. Increases in neutrophil levels of cyclic GMP but not of cyclic AMP were associated also with the discharge of ß-glucuronidase provoked by particles in the absence of added cholinergic agents. The data suggest that the immunologic release of lysosomal enzymes from human neutrophils can be regulated by autonomic neurohormones, perhaps via the selective formation of appropriate nucleotides.
EFFECTOR CELL BLOCKADE
Authors: Schrader John W. , Nossal G. J. V.
Published: June 2004

This study describes the effects of incubating antibody-forming cells (AFC), either as mass cell suspensions, or as single AFC in microdroplets, with antigens against which the cells display specificity. Most of the work was done with hapten-specific anti-DNP-AFC, but AFC with specificity against flagellar antigens or fowl gamma globulin (FGG) were also included. It was noted that 30-min incubation of AFC with highly multivalent forms of antigen caused a substantial partial suppression of the antibody-forming performance of the AFC as measured by a hemolytic plaque test. Thus, when cell suspensions containing anti-DNP plaque-forming cells (PFC), were incubated for 30 min at 37°C with 100 µg of DNP-polymerized flagellin (DNP-POL), the number of plaques appearing after washing of the cells and placing them in plaque-revealing erythrocyte monolayers was reduced to 50% or less compared with the number of plaques observed with control portions preincubated with medium alone. Preincubation with DNP-lysine, with oligovalent DNP-protein conjugates, or with irrelevant antigens produced no such inhibition. Studies where preinhibited PFC suspensions were mixed with control suspensions before assay showed that a nonspecific carryover of antigen into the assay system was not involved. The inhibitory effect could also be initiated by holding cells at 0°C with DNP-POL, but in that case, inhibition only became manifest after cells were incubated for 30 min at 37°C before being placed in plaque-revealing monolayers. This suggested that inhibition was initiated by adsorption of multivalent antigen onto PFC-surface Ig, but required some active process before secretion actually slowed down. The effect was dose- and time-dependent, antigen-specific, and generalized for all antigens studied.

As well as yielding reduced plaque numbers, the preinhibited cells also gave smaller, more turbid plaques, suggesting a reduction in antibody-forming rate by each PFC rather than the elimination of PFC. Consistent with this suggestion was the observation that the degree of inhibition of plaque formation could be increased by decreasing the sensitivity of the assay so that only AFC secreting at high rates were detected. A micromanipulation study, where single PFC were subjected to inhibition, and were then tested for the rate at which they could cause hemolysis, showed a 68% inhibition of mean secretory rate.

Micromanipulation studies were performed to test the amount of cell surface-associated Ig on control and preinhibited PFC. For this, single PFC were held with [125I]antiglobulin and quantitative radioautography was performed. No significant difference emerged, suggesting that retention of secreted Ig on cell-attached antigen was not the cause of inhibition.

The results are discussed in the framework of tolerance models and blocking effects at the T-cell level by antigen-antibody complexes. The name effector cell blockade is suggested in the belief that the phenomenon may be a general one applying to both T and B cells.
ROLE OF COMPLEMENT IN INDUCTION OF ANTIBODY PRODUCTION IN VIVO
Authors: Pepys M. B.
Published: June 2004

In an in vivo study in mice, suppression by the C3-cleaving protein of cobra venom (CoF), and other C3-reactive agents (zymosan, aggregated IgG, anti-C3 antibodies, and type III pneumococcal polysaccharide) of the thymus-dependent antibody responses to sheep erythrocytes, ovalbumin, and human IgG was demonstrated. The thymus-independent antibody response to polyvinyl-pyrrolidone was however unaffected by CoF.

These and other published observations suggest that there may be a requirement for functional C3 in induction of thymus-dependent but not thymus-independent antibody production. A model for the role of C3 in lymphocyte cooperation is proposed based on these data analyzed in the light of existing knowledge of this process. It is postulated that fixed C3 interacting with macrophage

See PDF for Structure

and B-cell C3 receptors might enhance or facilitate T-dependent presentation of antigen to B cells.
ACTIVATION OF T AND B LYMPHOCYTES IN VITRO
Authors: Armerding Dieter , Katz David H.
Published: June 2004

The studies presented herein have focused on the biological and biochemical properties of a nonspecific mediator produced by populations of activated T lymphocytes during short-term in vitro reactions with foreign alloantigens. We have analyzed the activity of the unseparated and of chromatographically separated fractions of the supernatants from such cultures on the in vitro responses of mouse lymphocytes to soluble and macrophage-bound DNP-carrier conjugates and also to particulate heterologous erythrocytes. The results demonstrate that a highly active protein moiety, termed allogeneic effect factors (AEF), in the mol wt range of 30,000–40,000, is capable of acting directly on B lymphocytes, in the presence of antigen, probably to effect triggering and subsequent differentiation and proliferation to antibody-forming cells in vitro. The active molecule, although not manifesting specificity for antigen, does exhibit some strain-specific properties suggesting a possible relationship to cell surface antigens or other gene products coded in the major histocompatibility gene complex.
AN OUTER MEMBRANE PROTEIN OF NEISSERIA MENINGITIDIS GROUP B RESPONSIBLE FOR SEROTYPE SPECIFICITY
Authors: Frasch Carl E. , Gotschlich Emil C.
Published: June 2004

Meningococcal groups B and C have been subdivided into a series of serotypes based upon the antigenic specificity of protein serotype antigens (STA). The purpose of these studies was to obtain the STA by gentle methods and determine its anatomic location in the meningococcal cell. The STA was extracted from group B meningococcal strains by either 0.2 M LiCl or 0.2 M CaCl2 and isolated from the extracts by gel filtration on Sepharose 6B or by pelleting the STA by centrifugation at 100,000 g. The isolated STA was a lipoprotein-lipopolysaccharide complex with a mol wt of approximately 4 x 106 daltons. Antisera prepared against the type 2 STA were bactericidal only for homologous serotype strains. The STA proved to be a constituent of the outer membrane of the cell envelope. This was shown by SDS-polyacrylamide gel electrophoresis (PAGE) of the isolated outer membrane and of the purified STA. The type 2 STA complex contains three principal proteins, one of which is predominant with a mol wt of 41,000 daltons. The type 2 STA was dissociated by Triton X-100 and separated by sucrose gradient isodensity centrifugation into two peaks. The denser peak (ρ = 1.26 g/cm3) contained the majority of the 41,000 dalton major outer membrane protein as shown by SDS-PAGE. This peak also contained the type 2 antigenic determinant. Thus the major outer membrane protein, extracted as part of a lipoprotein-lipopolysaccharide complex, contains the type 2 STA determinant.
RENAL DEPOSITION OF SOLUBLE IMMUNE COMPLEXES IN MICE BEARING B-16 MELANOMA
Authors: Poskitt Paula K. F. , Poskitt Thomas R. , Wallace John H.
Published: June 2004

Histologic and immunofluorescence studies of the kidneys of mice bearing a progressive melanoma show a proliferative glomerulonephritis associated with immune complex deposition in the mesangium and along the glomerular basement membrane This immune complex disease is distinct from the age-associated disease of the C57BL/6J host strain and the complexes can be shown to consist of soluble tumor antigen and antitumor antibody. Furthermore, the intensity of IgG complex deposition correlates directly with tumor progress (size and metastases) and inversely with mononuclear leukocyte infiltration of the tumor. In vitro assays for lymphocyte cytotoxicity and humoral antibody were found to be less reliable indicators of tumor progress. The possible role of circulating soluble tumor antigen in modifying the immune response to tumors is discussed.
ALLOANTISERUM-MEDIATED SUPPRESSION OF HISTOCOMPATIBILITY-LINKED Ir-GENE-CONTROLLED IMMUNE RESPONSES
Authors: Bluestein Harry G.
Published: June 2004

Fab, Fc, and F(ab)'2 fragments were prepared by enzymatic hydrolysis of the IgG fraction of strain 13 antistrain 2 alloantisera. These fragments were not cytotoxic to lymphocytes bearing strain 2 histocompatibility antigens, but the Fab and F(ab)'2 fragments retained functional combining sites as indicated by their ability to suppress the cytotoxicity mediated by the intact antistrain 2 antibodies. The F(ab)'2 fragments were much more efficient as inhibitors in this system than the Fab fragments. F(ab)'2 at 0.06 mg/ml and 0.45 mg/ml Fab produced comparable degrees of suppression. The F(ab)'2 at 0.06 mg/ml completely suppressed DNP copolymer of L-glutamic acid and L-lysine (GL)-stimulated tritiated thymidine incorporation. The monovalent Fab at 0.45 mg/ml, however, had no significant effect on the in vitro responses to DNP-GL. Addition of the intact alloantisera can be delayed 3 h after initiation of the antigen-stimulated cultures with no loss of suppression. After a delay of 6 h 45% suppression was observed. The requirement for the divalent molecule and the observation that effective suppression of the in vitro responses is still obtained when the alloantiserum is added several hours after initiation of the cultures both suggest that the immunosuppression results from an active process affecting the lymphocyte membrane that renders the cell refractory to the antigenic stimulus.
CELL-BOUND RECEPTORS FOR ALLOANTIGENS ON NORMAL LYMPHOCYTES
Authors: Binz Hans , Lindenmann Jean , Wigzell Hans
Published: June 2004

The rat popliteal node graft-vs.-host assay was shown to depend on the presence of parental T cells in the inoculum. Antialloantisera raised in F1 hybrid rats against alloantibodies of one parent directed at transplantation antigens of the other parent displayed some or all of the following specific effects on parental T cells: They inhibited local GvH by purified T-cell suspensions; they blocked the capacity of GvH-reactive cells to adsorb onto fibroblast monolayers of the relevant genotype; together with complement, they killed GvH-reactive cells.
EVIDENCE FOR IDENTITY OR CLOSE ASSOCIATION OF THE Fc RECEPTOR OF B LYMPHOCYTES AND ALLOANTIGENS DETERMINED BY THE Ir REGION OF THE H-2 COMPLEX
Authors: Dickler Howard B. , Sachs David H.
Published: June 2004

Immunoglobulin complexes, composed of heat-aggregated human Ig, were shown to bind to mouse B lymphocytes of a variety of strains, but not to either thymocytes or thymus-derived (T) lymphocytes under a variety of conditions. It was shown that this binding was not due to either natural human antibodies against mouse nor to nonspecific binding of human Ig by mouse lymphocytes. Such complexes were shown to bind to the same sites which bind mouse antibody-antigen complexes. This site is known as the Fc receptor.

The binding of Ig complexes to mouse B lymphocytes was markedly inhibited by pretreatment of the lymphocytes with anti-H-2 antisera. A series of experiments indicated the specificity of this result, including the fact that this inhibition was shown not to be due to the artifact of shedding of H-2 antibody-antigen complexes, nor to nonspecific steric inhibition.

The antibodies within anti-H-2 antisera which were responsible for this inhibition were specific for alloantigens associated with the Ir region of the H-2 complex (Ia antigens). Antiserum specific for these Ia antigens produced inhibition, whereas antisera specific for antigens determined by the K or D regions of the H-2 complex did not. Evidence was obtained using F1 hybrid cells that at least some Ia antigens of both parental types are expressed on every B lymphocyte (i.e. codominant expression).

These data indicate that the Fc receptor and a series of alloantigens controlled by the Ir region of the H-2 complex are identical or closely associated on the B-lymphocyte surface membrane. This observation may have implications for the mechanism of control of the immune response.
DIRECT EVIDENCE FOR HAGEMAN FACTOR (FACTOR XII) ACTIVATION BY BACTERIAL LIPOPOLYSACCHARIDES (ENDOTOXINS)
Authors: Morrison David C. , Cochrane Charles G.
Published: June 2004

Purified precursor Hageman factor has been demonstrated to bind to soluble bacterial lipopolysaccharide (LPS, endotoxin) isolated from Escherichia coli 0111:B4, and this complex has been shown to have the capacity to convert prekallikrein to its active form. In addition, LPS-activated Hageman factor substantially reduces clotting times in XII-deficient plasma. The capacity to activate Hageman factor has been demonstrated to reside in the lipid A region of the LPS molecule. Activation of Hageman factor by LPS contrasts with fluid-phase activation (e.g., by kallikrein or trypsin) in that no cleavage to lower molecular weight fragments occurs. High concentrations of LPS inhibit the activity of Hageman factor, probably by a direct LPS-Hageman factor interaction.