bluuebunny/crossref_metadata_2025 huggingface dataset:
Chemiluminescence of phagocytic cells caused by N-formylmethionyl peptides
Authors: Hatch GE , Gardner DE , Menzel DB
Published: June 2004

N-formylmethionyl (F-Met) peptides, when added alone to macrophages or polymorphonuclear leukocytes (PMN), were found to induce a chemiluminescent response of shorter duration than that produced by the commonly employed particulate stimulant, zymosan. The cellular nature of F-Met peptide-induced chemiluminescence was indicated by its dependence on cell concentration, and by its inhibition by cell disruption, heat inactivation, or previous maximal stimulation by the peptides.

Comparison of PMN and macrophages from different species showed that the maximal chemiluminescent response seen in the dose-response curve of F-Met- Phe was different in different cell types. Chemiluminescence reached highest values in human PMN, it was intermediate in guinea pig macrophages and PMN, and in rabbit PMN; but it was nonexistent in rabbit alveolar macrophages and very low in rabbit peritoneal macrophages. A definite relationship was observed between peptide structure and chemiluminescent activity. Met-Phe, F- Met and Phe were inactive even at millimolar concentrations, while F-Met-Phe caused chemiluminescence at micromolar concentrations. Four active peptides were tested in guinea pig, rabbit, and human PMN, and in guinea pig alveolar and peritoneal macrophages. The relative activity of these peptides was the same in all cells studied, e.g. F-Met-Leu-Phe >> F-Met-Phe > F-Met-Val > F- Met-Ala. The values of ED50 for each peptide were also comparable to previously reported ED50 values of these peptides in inducing lysosomal enzyme release. These results were seen both in the presence and absence ofthe chemiluminescent oxidant indicator, luminol.

Low concentrations of superoxide dismutase (10 μg/ml) completely inhibited chemiluminescence caused by the F-Met peptides, suggesting the involvement of 0(2)(-) or O(2)(-)-derived compounds in this response. Sodium azide, an inhibitor of peroxidase reactions, had either no effect or a slight inhibitory effect on chemiluminescence. However, when the extracellular release of lysosomal enzymes was induced by cytochalasin B, an azide- inhibitable enhancement of chemiluminescence was seen in PMN, but not in macrophages. This effect appears to be correlated with the presence of granule-associated myeloperoxidase. Although azide-inhibitable peroxidases could be a potential source of light, they did not appear to be a significant contributor in these experiments.

Based on these results and on those of previous investigators, we postulate that the F-Met-peptides stimulate 0(2)(-) production in addition to stimulating lysosomal enzyme release and chemotaxis. The similar structure- activity relationship which appears to exist for these processes may indicate that they are all initiated by a single receptor mechanism. Since F-Met peptides are formed in bacteria it is likely that their actions represent an important physiologic response.
Ontogeny of B-lymphocyte function. V. Thymus cell involvement in the functional maturation of B-lymphocytes from fetal mice transferred into adult irradiated hosts.
Authors: Sherr D H , Szewczuk M R , Siskind G W
Published: June 2004

Lethally irradiated mice reconstituted with adult thymus cells and neonatal or fetal liver cells produce an anti-2,4-dinitrophenyl or anti-bovine gamma globulin response of restricted heterogeneity of affinity in comparison with the response of mice reconstituted with B cells from adult donors. In addition, mice reconstituted with day 15 fetal B cells and adult thymus cells produce relatively few indirect plaque-forming cells (PFC). It was found that B cells acquire the capacity to produce a heterogeneous response, of predominantly indirect PFC within 7 days of transfer only when thymus cells are transferred along with the B cells. B cells from fetal or neonatal donors transferred without young adult thymus cells develop the capacity to generate indirect PFC within 13 days after transfer to adult recipients, but continue to produce a response of restricted heterogeneity of affinity for up to 28 days after transfer. Thus, it has been shown that cells present in the thymus facilitate, or are necessary for the functional maturation of B lymphocytes. Furthermore, the data suggest that maturation of the B-cell population to produce a heterogeneous response is controlled independently of its maturation to be capable of producing indirect PFC.
T-cell-mediated cytotoxic immune responses to F9 teratocarcinoma cells: cytolytic effector T cells lyse H-2-negative F9 cells and syngeneic spermatogonia.
Authors: Wagner H , Starzinski-Powitz A , Röllinghoff M , Golstein P , Jakob H
Published: June 2004

Murine thymus derived (T) lymphocytes primed in vivo to mouse 129 (H-2bc) derived H-2-negative F9 embryonal carcinoma cells and rechallenged in vitro with X-irradiated F9 stimulator cells differentiated into anti-F9 cell immune cytotoxic T lymphocytes (CTL). Using CBA mouse derived splenic responder T cells, F9 stimulator cells triggered a primary cytotoxic anti-F9 response. The CTL generated lysed the F9 antigen-positive target cells F9. PCC3 and PCC4, but not the F9 antigen-negative mouse 129 derived PYS tumor cells, nor LPS induced H-2bc blast cells. Mouse 129 anti-F9 cell antisera but not H-2k anti-H-2bc antisera blocked the lytic interaction with F9 target cells. Similarily unlabeled F9 cells but not H-2bc blast cells inhibited the anti-F9 cell cytotoxicity H-2k anti-F9 cell immune CTL were found to be cytotoxic for syngeneic spermatogonia, known to express the F9 antigen. The results suggest not only that CTL can recognize and lyse H-2-negative target cells, but also that CTL precursors can be sensitized against H-2-negative stimulator cells. From the data available it may be inferred that anti-F9 Cell immune CTL recognize the F9 antigen, known to be linked with the T/t locus. Since anti-F9 cell immune CTL lyse syngeneic spermatogonia, the system may be useful to analyze in vitro the induction and effector phase of a T-cell-mediated cytotoxic autoimmune orchitis.
Nonrandom inclusion of H-2K and H-2D antigens in Friend virus particles from mice of various strains.
Authors: Bubbers J E , Chen S , Lilly F
Published: June 2004

Friend murine leukemia virus (FV), isolated from infectious serum of several mouse strains, has been examined for the presence of H-2 antigens. Following banding of the virus on a discontinuous sucrose gradient, pelleting, and disruption with Nonidet P-40 detergent, virus preparations were tested for their capacity to inhibit the lysis of target cells mediated by various anti-H-2K or anti-H-2D antisera. Virus from mice homozygous for the H-2b, H-2d, H-2g H-2k, and H-2ol haplotypes or heterozygous for the H-2g/H-2k, H-2b/H-2d, and H-2b/H-2k haplotypes was used. Of the six H-2D or H-2K alleles examined, the products of only two, H-2Db and H-2Kk were detected. Virus preparations contained no, one, or both antigens, depending on the genotype of the host animal. Control preparations from normal mouse serum and preparations in which the virus had not been disrupted demonstrated no H-2 activity. Furthermore, attempts to neutralize FV spleen focus forming activity with anti-H-2Db or anti-H-2Kk antisera yielded negative results.
Genetic control of the immune response to staphylococcal nuclease. VII. Role of non-H-2-linked genes in the control of the anti-nuclease antibody response
Authors: Pisetsky DS , Berzofsky JA , Sachs DH
Published: June 2004

The role of non-H-2-linked genes in the control of the antibody response to staphylococcal nuclease has been investigated. 3 wk after immunization with nuclease in complete Freund's adjuvant, strain A/J (H-2 a) mice produced significantly higher titers of antibody than strain B10.A (H-2(a)) mice, whereas mice of strains A.BY (H-2(b)) and B10 (H-2(b)) produced barely detectable titers. With hyperimmunization, A/J and A.BY mice reached the same peak levels for antibody titers, both severalfold higher than those reached by B10.A and B10 mice. Analysis of the specificity of antibodies by assessment of binding to two fragments of nuclease showed similarities between strains of the same H-2 haplotype. These results suggest that although H-2-1inked genes determined initial responsiveness at 3 wk and the relative proportions of antibodies directed toward different antigenic determinants on the nuclease molecule, non-H-2-linked genes determined the overall magnitude of the hyperimmuneresponse. Measurement of the affinity of the antibodies to the nuclease fragment (1-126) showed that strains B10 and B10.A produced antibodies with 7- to 10-fold higher affinity than comparable antibodies from strains A.BY and A/J. In a backcross of (B10.A × A/J) × B10.A, the level of antibody segregated independently of the Ig-1(e) C(H) allotype and the A/J anti-nuclease idiotypes. Thus, a gene(s) linked to neither H-2 nor heavy chain structural genes appears to control the aggregate response to antigenic determinants on the nuclease molecule independent of subspecificities of these antibodies or their idiotype.
The genetic basis of susceptibility to leukemia induction in mice by 3-methylcholanthrene applied percutaneously.
Authors: Duran-Reynals M L , Lilly F , Bosch A , Blank K J
Published: June 2004

Susceptibility to leukemia induction in mice by skin painting with 3-methylcholanthrene (MCA) is strain-specific, occurring only in strains relatively resistant to MCA-induced skin tumors. The Ah locus, which has a dominant allele (Ahb) for inducibility of the aryl hydrocarbon hydroxylase (AHH) enzyme system and a recessive allele (Ahd) for noninducibility, appears to be the major determinant of this trait. MCA-painted mice of strains and crosses carrying the Ahb allele usually show a high incidence of skin tumors (papillomas which may evolve into malignant tumors) and little or no leukemia, whereas in mice homozygous for the Ahd allele the treatment usually induces a high incidence of leukemia and few or no skin tumors. Among mice of a segregating backcross generation including both Ahb/Ahd heterozygotes and Ahd homozygotes, the occurrence of skin tumors was correlated directly with AHH inducibility and inversely with the leukemic response. Mice of Ahb strains with a high level of endogenous murine leukemia (MuLV) expression (C58, PL) show a much weaker skin tumor response than expected but no increase in leukemia incidence, and this observation tends to confirm the previous finding that MuLV infection of mice of low-MuLV strains results in reduced susceptibility to MCA tumorigenesis.
Characterization of human high molecular weight kininogen. Procoagulant activity associated with the light chain of kinin-free high molecular weight kininogen.
Authors: Thompson R E , Mandle R , Kaplan A P
Published: June 2004

Human high molecular weight (HMW) kininogen has been isolated and was found to be a single chain protein of approximately equal to 120,000 daltons. Upon digestion with plasma kallikrein bradykinin is generated, and SDS gel electrophoresis of the kinin-free protein reveals an apparent loss in size of 15,000 daltons. The kinin-free kininogen retains full activity as a coagulation factor and consists of two chains: a heavy chain of approximately equal to 66,000 daltons disulfide-linked to a light chain of 37,000 daltons. The heavy chain of HMW kininogen shares antigenic determinants with LMW kininogen and possesses no detectable coagulant activity. The isolated light chain is shown to be responsible for the coagulant activity of HMW kininogen and contains a unique antigenic determinant that distinguishes HMW kininogen from low molecular weight kininogen.
The role of H-2 linked genes in helper T-cell function. II. Isolation on antigen-pulsed macrophages of two separate populations of F1 helper T cells each specific for antigen and one set of parental H-2 products.
Authors: Swierkosz J E , Rock K , Marrack P , Kappler J W
Published: June 2004

A method was established for isolating antigen-specific murine helper T cells by selective binding to antigen-pulsed macrophage (Mphi) monolayers. Sheep erythrocyte (SRBC)-primed T cells, which remained strongly adherent to SRBC-pulsed syngeneic Mphi after 20 h in culture, were markedly enriched for helper activity when tested in the in vitro antitrinitrophenol (TNP) response to TNP-SRBC. Successful binding and enrichment occurred only if the Mphi were pulsed with the specific antigen to which the T-cell donors had been primed. The genetic control governing helper function in this system was then examined by using primed F1 T cells isolated on Mphi monolayers from congenic strains bearing parental H-2 haplotypes. SRBC-primed BDF1 (H-2b X H-2d) T cells, which bound to SRBC-pulsed H-2d Mphi, subsequently functioned as helper cells in cultures containing H-2d B cells and Mphi, but not in those containing H-2b B cells and Mphi. They remained unable to collaborate with B cells of the H-2B haplotype even in the presence of additional H-2d Mphi, indicating that H-2 restriction occurs at least at the level of the B cell. Similary, primed BDF1 T cells isolated on H-2b Mphi cooperated preferentially with H-2b B cells and Mphi. In both cases, the haplotype preference of the T cell was not due to alloreactive suppressor activity. These results suggest that primed F1 mice contain individual populations of helper T cells, each of which recognize antigen in association with a parental H-2 gene product(s) expressed during both Mphi-T cell and T cell-B cell interactions.
Macrophage activation: increased ingestion of IgG-coated erythrocytes after administration of interferon inducers to mice.
Authors: Hamburg S I , Manejias R E , Rabinovitch M
Published: June 2004

In vitro phagocytosis of IgG-opsonized sheep erythrocytes (EA) was used to measure the in vivo activation of mouse peritoneal macrophages. Uptake of EA as enhanced by the extraperitoneal administration of Newcastle disease virus, vesicular stomatitis virus, tilorone or polyinosinic-polycytidylic acid. Ingestion of EA was similarly stimulated by lipopolysaccharide or killed Corynebacterium parvum. Dose-response curves relating concentrations of IgG to phagocytosis were parallel for both treated and control animals. This indicates that the heterogeneity of the macrophage populations did not change and that the overall populations were activated with respect to phagocytic ability. Numbers of macrophages were not increased (except in C. parvum-treated mice), suggesting that resident, rather than newly recruited macrophages, were activated by the different agents.
Expression of a single major histocompatibility complex locus controls the immune complex locus controls the immune response to poly-L-(tyrosine, glutamic acid)-poly-DL-alanine--poly-L-lysine
Authors: Deak BD , Meruelo D , McDevitt HO
Published: June 2004

Genetic control of the immune response linked to the major histocompatibility (H-2) complex in the mouse has been described for synthetic polypeptide antigens and for low doses of native proteins. The phenomenon is well documented(1,2). Extensive screening of intra-H-2 crossover-derived recombinant strains has localized H-2-linked immune response (Ir) genes to the I-immune response region of the H-2 complex (3). For most antigens, Ir genes are autosomal, dominant, and they segregate as single loci. It is not known whether these crossover-defined loci respresent single genes with multiple alleles or clusters of tightly linked genes (4).

In 1972, Stimpfling and Durham (5) postulated that two interacting loci within the H-2 complex were required for the response to the alloantigen, H-2.2 (6), and, in 1975, Dorf et. al. (7) observed a responder phenotype in a recombinant derived from two strains which were nonresponders to the synthetic linear terpolymer, L-glutamic acid, L-lysine, L-phenylaline (GLPhe). Analysis of additional recombinants and complementation tests with F(1) hybrids clearly demonstrated that genes in two intra-I-region loci controlled the immune response to GLPhe. Subsequently, requirement for genes mapping in two intra-I-region loci were reported for porcine LDH(B)(8), the alloantigen Thy-1.1 (9), and for the synthetic terpolymers L-glutamic acid, L-lysine, L-tyrosine and L-glutamic acid, L-lysine, L- leucine (6,10).

Demonstration that responses to both synthetic polypeptide and native protein antigens can be controlled by genes in two distinct I-region loci prompted speculation that the phenotypic expression of two I-region genes is a general phenomenon which may provide the key for understanding the mechanism of Ir gene function and cellular collaboration in the immune response. Benacerraf and Dorf (10) have shown that Ir gene complementation is often more effective in the cis than in the trans configuration. This concept is further supported by the data reported for GLPhe (10-12) which indicate that both of the complementing genes must be expressed in each of the cell types participating in the interaction. Failure to detect complementation for the majority of antigens under H-2-linked Ir-gene control might be attributed to the limited number of available intra-I- region recombinant strains.
The differential destructive and enhancing effects of anti-H-2K, H-2D, and anti-Ia antisera on murine skin allografts.
Authors: McKenzie I F , Henning M M
Published: June 2004

Nude (athymic) or anti-lymphocyte serum-treated mice have absent delayed graft rejection due to impaired T-cell responses. Nonetheless, these mice can reject skin grafts, acutely, when treated with anti-H-2 antibody and additional complement. Resolution of the components in the H-2 antisera, by either absorption or by selective production of antisera of restricted specificity has demonstrated that anti-H-2K or H-2D antiser are graft destructive, and as shown elsewhere, are nonenhancing. By contrast, anti-Ia sera are not capable of causing allograft destruction even when used in extremely high doses and were previously noted to be enhancing. The mechanism of such differential effects is not apparent, but the findings are clearly of importance to transplantation in man, where sera reacting specifically with B cells may also be enhancing and nondestructive.
Cytotoxic T-cell responses to H-Y: correlation with the rejection of syngeneic male skin grafts.
Authors: Hurme M , Chandler P R , Hetherington C M , Simpson E
Published: June 2004

The ability of female mice to rapidly reject syngenic male skin grafts is largely determined by dominant genes in the IB region of the H-2b halotype, whereas the ability to produce anti-H-Y cytotoxic cells is determined by a dominant gene in the IA region the H-2b halotype, or by complementary genes in the IC region of some other haplotypes. Thus, it seems that H-2-retricted anti-H-Y cytotoxic T cells are not responsible for the rejection of syngeneic male skin grafts.
Induction of partial chimerism in nonirradiated B-lymphocyte-deficient CBA/N mice.
Authors: Volf D , Sensenbrenner L L , Sharkis S J , Elfenbein G J , Scher I
Published: June 2004

Nonirradiated B-lymphocyte-deficient CBA/N mice given T6T6 chromosome-marked normal CBA/CaHN spleen cells became lymphoid chimeras exhibiting donor-type mitoses. Normal CBA/CaHN recipients did not exhibit significant numbers of donor-type mitoses. The lymphoid cell chimerism in the CBA/N host appeared in spleen, lymph nodes, and Peyer's patches, but not in marrow or thymus. Stimulation of CBA/N-recipient spleen cells in vitro suggested that the chimerism involved donor T6T6 cells which were responsive to the B-lymphocyte mitogen, lipopolysaccharide, but not to the T-lymphocyte mitogen, phytohemagglutinin. These data indicate that stable, long-term chimerism of a specific class of lymphocytes is possible in nonirradiated, B-lymphocyte-deficient CBA/N mice.
Suppression of the mixed lymphocyte reaction in man by a soluble T-cell factor. Specificity of the factor for both responder and stimulator.
Authors: Engleman E G , McMichael A J , McDevitt H O
Published: June 2004

J.H., an HLA-Dw2 homozygous multiparous woman, fails to respond to her husband, W.H. (HLA Dw1,-) in the unidirectional mixed lymphocyte reaction. T cells from J.H. were previously shown to suppress the responses of Dw2-positive cells but not Dw2-negative cells to W.H. We now report that a soluble factor released into the supernate of the mixed lymphocyte reaction by J.H. T cells, mediates this suppression. Like the cell from which it is derived, the factor is highly specific for HLA Dw2 in the responder cell and partially specific for the stimulatory alloantigen.
Distribution of a major connective tissue protein, fibronectin, in normal human tissues.
Authors: Stenman S , Vaheri A
Published: June 2004

Fibronectin is a major surface-associated glycoprotein of cultured fibroblasts and it is also present in human plasma. Antiserum specific for human fibronectin was used to study the distribution of fibronectin in normal adult human tissues. The protein was detected (a) characteristically in various basement membranes including capillary walls: (b) around individual smooth muscle cells and in the sarcolemma of striated muscle fibers; and (c) in the stroma of lymphatic tissue and as thin fibers in loose connective tissue. The distribution of fibronectin was distinct from that of collagen and elastic fibers, but was very similar to reticulin, as demonstrated by conventional histological staining. The results indicate that fibronectin is a major component of connective tissue matrix. The distribution also indicates that most types of adherent cells abut fibronectin-containing structures. This supports the possible role of fibronectin in cell-cell and cell-matrix interactions in tissues.
T cells specific for hapten-modified self are precommitted for self major histocompatibility complex antigens before encounter with the hapten.
Authors: Janeway C A , Murphy P D , Kemp J , Wigzell H
Published: June 2004

The technique of antigen-driven, 5-bromo-deoxyuridine and light suicide has been adapted to eliminate the precursors of cytotoxic effector cells both for alloantigen and for 2,4,6-trinitrophenyl(TNP)-modified stimulator and target cells. Using this technique, the following observations have been made. Precursors of killer cells specific for alloantigen can be suicided independently of precursors of killer cells specific for TNP-modified self cells. The loss of activity during this procedure is not due to either specific or nonspecific suppressor cells, as judged by mixing experiments. With responder cells from F1 animals, it has been possible to show that precursors specific for TNP-modified cells from one parent are suicided independently of precursors specific for TNP-modified cells of the other parent, but only if the parental strains differ in the K and D regions of the H-2 complex. Cells of F1 mice derived from K and D identical, I region different, parental strains were specifically suicided by TNP-modified stimulator cells from either parent. However, the cross-reactive killing of TNP-self targets induced by stimulation with allogeneic cells is not eliminated by first suiciding with TNP-parental cells, suggesting that the precursors of these two types of TNP-self killer cells are different. This is compatible with reported differences in their specificity, as confirmed in this report. Finally, deletion of alloreactive cells by this technique reveals little or no reactivity specific for TNP-modified allogeneic stimulator cells. In summary, these results strongly suggest that recognition of self MHC antigens is preprogrammed in peripheral T cells of normal animals, and is not acquired during the immunization process. They also suggest that cells specific for modified alloantigen are relatively rare in the strains of mice studied.
Restricted helper function of F1 hybrid T cells positively selected to heterologous erythrocytes in irradiated parental strain mice. I. Failure to collaborate with B cells of the opposite parental strain not associated with active suppression.
Authors: Sprent J
Published: June 2004

Unprimed (CBA X C57BL/6)F1 lymph node T cells were transferred with sheep erythrocytes (SRC) into heavily irradiated F1 or parental strain mice and recovered from thoracic duct lymph or spleens of the recipients 5 days later. To study their helper function, the harvested F1 T cells were transferred with antigen into irradiated F1 mice plus B cells from either the two parental strains or from F1 mice. F1 T cells activated in F1 mice gave high IgM and IgG anti-SRC responses with all three populations of B cells. By contrast, F1 T cells activated in mice of one parental strain collaborated well with B cells of this strain, but poorly with B cells of the opposite strain. Active suppression was considered an unlikely explanation for this result since (a) good responses were found with F1 B cells, and (b) addition experiments showed that the poor response with B cells of the opposite parental strain (which was equivalent to that produced by unprimed F1 T cells) could be converted to a high response by a supplemental injection of F1 T cells activated in F1 mice. The phenomenon (a) was specific for the antigen used for activation (criss-cross experiments were performed with horse erythrocytes), (b) was reflected in levels of serum hemagglutinins as well as in numbers of splenic plaque-forming cells, (c) applied also to comparable activation of (DBA/2 X C57BL/6)F1 T cells, and (d) could be prevented by activating F1 T cells in mice of one parental strain in the presence of peritoneal exudate cells of the opposite parental strain. The hypothesis was advanced that F1 T cells contain two discrete subpopulations of antigen-reactive cells, each subject to restrictions acting at two different levels: (a) during T-macrophage interactions and (b) during T-B collaboration. It was suggested that when F1 T cells are activated to antigen in a parental strain environment, radioresistant macrophages activate only one of the two subgroups of T cells, and this subgroup is able to collaborate with B cells of the strain used for activation (and with F1 B cells) but not with B cells of the opposite parental strain. The other subgroup of T cells remains in an unprimed (nonactivated) state.
Restricted helper function of F1 hybrid T cells positively selected to heterologous erythrocytes in irradiated parental strain mice. II. Evidence for restrictions affecting helper cell induction and T-B collaboration, both mapping to the K-end of the H-2 complex.
Authors: Sprent J
Published: June 2004

Studies with H-2-congenic and recombinant strains showed that when F1 hybrid T cells were activated to sheep erythrocytes in irradiated mice of parental strain or related strain, a population of helper cells was generated which collaborated only with B cells sharing the K-end of the H-2 complex with the strain used for activation. No evidence was found that the restriction in helper function (a) reflected a deficiency of appropriate macrophages during T-B collaboration, or (b) was influenced by the Ig allotype of the B cells. It was concluded that the results signified restrictions acting at both the level of helper cell induction (presumed to be a reflection of T-macrophage interactions in the irradiated intermediate hosts) and during T-B collaboration. With (CBA X C57BL/6)F1 cells, the restrictions at each level mapped to the same region i.e. to the left of the I-B subregion. Consequently, one gene (or set of genes) might control restriction at both levels. If so, T-cell recognition of major histocompatibility complex-associated antigen on macrophages and on specific B cells would be either identical or very similar. The fact that genes mapping to the K-end of the H-2 complex also control the restrictive interactions of homozygous T cells implies that F1 T cells behave functionally as a mixture of T cells derived from the two parental strains. Positive selection to antigen in parental strain mice appears simply to alter the ratio of these two populations.
Effect of colchicine on the antibody response. II. Demonstration of the inactivation of suppressor cell activities by colchicine.
Authors: Shek P N , Waltenbaugh C , Coons A H
Published: June 2004

The simultaneous administration of colchicine (CC) with a T-independent antigen, e.g. 2,4,6-trinitrophenyl-keyhold limpet hemocyanin-Sepharose, to intact animals effectively enhanced their hapten-specific plaque-forming cell (PFC) response. However, in congenitally athymic nude mice in which T-cell regulation was absent, CC was ineffective in producing enhancement. These observations suggest that the target cell acted upon by CC is most likely thymus-derived. Furthermore, the injection of CC with the co-polymer of L-glutamic acid50-L-tyrosine50 (GT) abolished GT-specific suppression of the PFC response to GT-methylated bovine serum albumin. Spleen cells from CC-treated and GT-primed hosts could no longer transfer suppressive activity to normal recipients. These results provide evidence that CC is capable of inactivating or eliminating suppressor cells or their precursors. Thus, CC-induced enhancement of the antibody response may be explained, at least in part, by its antimitotic, and hence lethal effect on dividing suppressor T cells.
Proliferative capacity of mouse peritoneal macrophages in vitro.
Authors: van der Zeijst B A , Stewart C C , Schlesinger S
Published: June 2004

Thioglycolate-stimulated mouse peritoneal macrophages cultured in the presence of macrophage growth factor (MGF) will continue to proliferate when they are removed from culture dishes with the local anesthetic lidocaine and subcultured. The number of times the cells can be subcultured and remain in a proliferative state is dependent on the number of previous cell divisions. One precursor cell (colony-forming cell) yields about 2.6 X 10(4) daughter cells. When MGF is removed from actively proliferating macrophages, they leave the cell cycle and enter a "resting" condition. When MGF is readded, cells reenter the cell cycle and proliferate with the same doubling time as if MGF had not been removed. Membrane 5'-nucleotidase activity was used as a probe to identify the state of macrophage activation. Proliferating macrophage populations had significantly higher enzyme levels than stimulated macrophages cultured without MGF. These enzymes levels were, however, lower than those found for resident (unstimulated) macrophages.
Isolated hapten-binding receptors of sensitized lymphocytes. III. Evidence for idiotypic restriction of T-cell receptors.
Authors: Krawinkel U , Cramer M , Melchers I , Imanishi-Kari T , Rajewsky K
Published: June 2004

The primary antibody response of C57BL/6 mice to the 4-hydroxy-3-nitrophenylacetyl (NP) hapten is restricted to antibody molecules expressing the NPb idiotype. This idiotype is a genetic marker for V genes in the heavy chain linkage group. In the secondary response, the frequency of NPb idiotype-positive molecules within the antibody population drops to very low values. Accordingly, isolated NP binding receptors from NP-sensitized B lymphocytes are largely devoid of this idiotype. In contrast, the NPb idiotype is expressed on the majority of the receptor fractions which we consider T-cell derived. This finding suggests that the antigen receptors of T lymphocytes may be restricted to the expression of major (germ-line encoded?) heavy chain idiotypes.
Primary in vitro cell-mediated lympholysis reaction of NZB mice against unmodified targets syngeneic at the major histocompatibility complex.
Authors: Botzenhardt U , Klein J , Ziff M
Published: June 2004

T-cell cytotoxicity of NZV mice was tested after in vitro sensitization against a group of H-2 identical strains (BALB/c, B10.D2, DBA/2, HW19). A highly significant and unexpected unidirectional cell-mediated lympholysis (CML) reaction by the sensitized NZB effector cells on these targets was found. After sensitization in vitro with stimulator cells of one H-2d strain, NZB effector cells (H-2d) lysed all other H-2d targets and to a lesser degree, some non-H-2d targets (C57BL/10, DBA/1, B10.Q, CBA, B10.S, A.SW). NZB targets were not lysed. Differences in the major histocompatibility region between NZB and other H-2d strains could be excluded as a possible explanation for the observed reaction of NZB (H-2d) against other H-2d strains. These results consequently represent the first description of a primary in vitro CML directed against determinants not coded for in the major histocompatibility complex. The responsible effector cells are demonstrated to be T cells. The CML of NZB against H-2 identiical targets appears best explained by a reaction against minor histocompatibility antigens. This, and the observed cross-reactions, would indicate that the cytotoxic T-cell system in NZB mice is not subjected to restrictions found in all normal mouse strains tested until now under similar conditions. It is suggested that this hyperreactivity is related to the autoimmune responsiveness of the NZB strain.
Genetic control of the immune response to staphylococcal nuclease. VIII. Mapping of genes for antibodies to different antigenic regions of nuclease.
Authors: Pisetsky D S , Sachs D H
Published: June 2004

Antibodies to staphylococcal nuclease have been fractionated into two populations on the basis of their ability to bind to the cyanogen bromide cleavage product of nuclease comprising the C-terminal portion of the molecule from the 99th to the 149th amino acid. The two populations of antibodies, anti-nuclease (1-99)n and anti-nuclease (99-149)N, have been prepared from a variety of strains, and analyzed using anti-idiotypic antisera raised against whole anti-nuclease antibodies from strains A/J, SJL, BALB/c, and B10.A(2R). Anti-nuclease (1-99)n, antibodies had the same pattern of reactivity with the anti-idiotypic antisera as did unfractionated antibodies, whereas a different pattern was found for anti-nuclease (99-149)n preparations. On the basis of these studies, five anti-nuclease idiotypes, designated NASE markers, have been identified and defined on the basis of their antigenic specificity and strain distribution. With these additional markers, it has been possible to provide more detailed maps of variable (V) region genes in the strains BALB/c, CB.20, and the recombinant BAB.14. A recombinational event between V region genes during the development of the BAB.14 strain is suggested by the positioning of these NASE markers.
Loss of proliferative capacity in immunohemopoietic stem cells caused by serial transplantation rather than aging.
Authors: Harrison D E , Astle C M , Delaittre J A
Published: June 2004

Marrow stem cell lines from old donors and those from young controls gave equally rapid rates of colony growth on spleens of irradiated mice. Old and young stem cell lines competed equally well with chromosomally marked marrow stem cells from a young donor in producing cell types that are stimulated by bleeding; old cells competed 70% as well as young in producing cell types stimulated by phytohemagglutinin (PHA) in vitro. After a single serial transplantation, the rates of colony growth declined 1.5- to 2.5-fold, and the ability to compete declined 2- to 4-fold for bleeding-stimulated and 4- to 10-fold for PHA-stimulated cells. Thus, immediate stem cell proliferative capacities decline much more after one serial transplantation than after a lifetime of normal function.
IgG2a-producing variants of an IgG2b-producing mouse myeloma cell line.
Authors: Francus T , Dharmgrongartama B , Campbell R , Scharff M D , Birshtein B K
Published: June 2004

12 variant cell lines producing an IgG2a (kappa) immunoglobulin derived via different routes from the IgG2b (kappa) synthesizing MPC 11 were studied. These variants all have the parental MPC 11 idiotype as shown by a radioimmunoassay. A comparison of the variants by charge, peptide maps, and assembly patterns has shown that most of them differ from one another, and some can be grouped. One group consists of three primary variants generated with two mutagenic agents: these three have almost indistinguishable peptide maps. Two other primary variants which arose in a similar fashion differ markedly from each other and from this group. A second group is comprised of the four secondary variants which arose from two short heavy chain producing primary variants. Other secondary variants and the one spontaneously arising variant cannot be grouped. Possible genetic mechanisms such as translocation, expression of previously silent genes and recombination are discussed.
Requirement for hexose, unrelated to energy provision, in T-cell-mediated cytolysis at the lethal hit stage.
Authors: MacLennan I C , Golstein P
Published: June 2004

The requirement for D-glucose in T-cell-mediated cytolysis was studied using mouse spleen cells sensitized against alloantigens in vitro. Glucose was required for cytolysis: (a) cytolysis proceeded in a simple buffered salt solution containing Ca++ and Mg++ (low phosphate-buffered saline, LPBS) in the presence but not in the absence of added glucose; (b) 2-deoxy-D-glucose blocked cytolysis. The block by this agent was overcome by excess glucose added as late as 40 min after the inhibitor. This block was not due to inhibition of NADP reduction, since 2-deoxy-D-glucose failed to interfere with the rate of CO2 production by the pentose cycle which we found to be of significant activity in sensitized spleen cells; (c) dialyzed fetal bovine serum (DFBS) in LPBS supported cytolysis in the absence of added glucose. However, 2-deoxy-D-glucose was also inhibitory under these conditions, suggesting that carbohydrate was required here as well. Further results supported the conclusion that DFBS was not acting as a direct source of the required carbohydrate. The relationship between cytolysis, glucose requirement, and provision of energy was studied. As little as 0.1 mM D-glucose in LPBS supported cytolysis. At this glucose concentration, there was no measurable accumulation of lactate in sensitized spleen cells, but Krebs cycle activity was detectable. In 3 mM glucose or above, the range covered by standard tissue culture media, anaerobic glycolysis became a major source of energy in sensitized spleen cells. Consequently, it appears that in standard tissue culture medium, effector cells can generate sufficient energy for cytolysis either by aerobic or anaerobic metabolism. However, the addition of an energy source alone in the absence of glucose was insufficient to support cytolysis in LPBS. Pyruvate in LPBS did not support cytolysis but was shown to be a good substrate for aerobic metabolism in sensitized spleen cells. Glycogenic amino acids and glycerol also failed to support cytolysis. The stage of cytolysis at which glucose is required was investigated. Glucose was necessary for the calcium-dependent lethal hit phase, but not for the cytochalasin A-blockable recognition stage, nor for 51Cr release from injured target cells. Models for the lethal hit process are discussed, which are compatible with the observed requirement for certain hexoses unrelated to their capacity to serve as sources of energy.
Effect of castration and sex hormone treatment on survival, anti-nucleic acid antibodies, and glomerulonephritis in NZB/NZW F1 mice.
Authors: Roubinian J R , Talal N , Greenspan J S , Goodman J R , Siiteri P K
Published: June 2004

NZB/NZW F1 mice of both sexes were castrated at 2 wk of age and implanted subcutaneously with silastic tubes containing either 5-alpha-dihydrotestosterone or estradiol-17-beta. Mice receiving androgen showed improved survival, reduced anti-nucleic acid antibodies, or less evidence of glomerulonephritis as determined by light, immunofluorescent, and electron microscopy. By contrast, opposite effects were observed in castrated mice receiving estrogen. Intact male NZB/NZW F1 mice received androgen implants at 8 mo, an age when they develop an accelerated autoimmune disease associated with a decline in serum testosterone concentration. Such treated mice had improved survival and reduced concentrations of antibodies to DNA and to polyadenylic acid (Poly A). Prepubertal castration of male NZB/NZW F1 mice results in an earlier appearance of IgG antibodies to Poly A. This effect of castration was prevented if neonatal thymectomy was also performed.
Cellular and genetic control of antibody responses in vitro. III. Immune response gene regulation of accessory cell function
Authors: Singer A , Cowing C , Hathcock KS , Dickler HB , Hodes RJ
Published: June 2004

The possibility was investigated that Ir genes regulate the function of cells other than T or B cells in the primary IgM responses to the synthetic antigens trinitrophenylated poly-L-(Tyr,Glu)-poly-D,L-Ala--poly-L-Lys [TNP-(T,G)-A--L]and trinitrophenylated poly-,-(His,Glu)-poly-D, L-Ala--poly-L-Lys [TNP-(H,G)-A--L]. The primary responses of (B10 × B10.A)F(1) spleen cells to both antigens were abrogated by Sephadex G-10 passage, and restored by the addition of spleen adherent cells. The cell type in the spleen adherent cell population active in reconstituting the responses to TNP-(T,G)-A--L and TNP-(H,G)-A--L was a non-T, non-B, radiation-resistant, glass-adherent spleen cell. The responses of Sephadex G-10-passed (responder x nonresponder)F(1) spleen cells to TNP-(T,G)-A--L or TNP-(H,G)-A--L were reconstituted by spleen adherent cells from only responder strains. Spleen adherent cells from F(1) mice reconstituted the responses to both antigens. Spleen adherent cells from each of the strains tested reconstituted the non- Ir gene-controlled response to a third antigen, TNP-keyhole limpet hemocyanin. The inability of spleen adherent cells from nonresponder strains to reconstitute the responses to either TNP-(T,G)-A--L or TNP-(H,G)-A--L was not a result of active suppression induced by the presence of nonresponder adherent cells, since a mixture of responder and nonresponder spleen adherent cells reconstituted the responses to both antigens. The use of spleen adherent cells from recombinant strains demonstrated that the autosomal dominant genes controlling the ability of spleen adherent cells to function as accessory cells in the responses to TNP-(T,G)-A--L and TNP-(H,G)-A--L are located in the K or I-A regions of the responder H-2 complex, the same region(s) of H-2 as the Ir genes controlling overall in vitro and in vivo responsiveness to these antigens.
In vitro induction of cytotoxic effector cells with spontaneous killer cell specificity.
Authors: Jondal M , Targan S
Published: June 2004

The present work shows that stimulation of peripheral blood lymphocytes with autologous or allogeneic B-cell lines leads to a strong induction of cytotoxic effector cells with spontaneous killer (SK) cell specificity, apart from the specific response directed against the particular stimulating cell. To demonstrate this we have determined a relative target cell specificity in the SK system, defined by the short-term 51Cr release assay, and established a relative specificity index (RSI). Using this approach we have been able to show that killer cells induced during a 5-day cocultivation period with B-cell lines have a similar PSI to that of unstimulated SK cells. In addition, we have shown that such killer cells can be induced from several different lymphocyte subpopulations and that they, in contrast to SK cells, do not express Fc receptors. The implications of these findings in relation to the nature, mechanism, and biological significance of the SK cell system is discussed.
Isolation of a heterogeneous population of temperature-sensitive mutants of measles virus from persistently infected human lymphoblastoid cell lines.
Authors: Ju G , Udem S , Rager-Zisman B , Bloom B R
Published: June 2004

Two human lymphoblastoid B-cell lines, WI-L2 and 8866, were infected with the Edmonston strain of measles virus at a multiplicity of infection of 10(-6), and stable persistent infections were established. By immunofluorescence and electron microscopy, the vast majority of cells from both cell lines were expressing viral antigens and releasing virion-like particles. However, very little infectious virus could be detected at 37 degrees C, either by an infectious centers assay or by titration of supernates from persistently infected cultures. When cultures were shifted to 31 degrees C, the cells released a population of virus that was temperature-sensitive. Clonal analysis of supernatant virus at 31 degrees C revealed a highly heterogeneous population of temperature-sensitive mutants, differing in plating efficiency ratios, thermolability, and antigen production at the nonpermissive temperature. Factors such as interferon, defective interfering particles, and extracellular virus do not appear to be important in maintaining the persistent carrier state. These studies have important implications for persistent infections of lymphoid cells in vivo, and the slow neurological diseases associated with measles, subacute sclerosing panencephalitis, and multiple sclerosis.
Human immune responses to hapten-conjugated cells. I. Primary and secondary proliferative responses in vitro.
Authors: Seldin M F , Rich R R
Published: June 2004

An in vitro model was developed to study both primary and secondary proliferative responses of human lymphocytes to hapten-conjugated peripheral blood mononuclear cells. Coculture of human lymphocytes with autologous trinitrophenyl (TNP)-conjugated stimulator cells resulted in primary proliferative responses. Subjects segregated into high and low primary responders with mean stimulation indices of 11 and 2.1, respectively. Restimulation of primed cells from high responder subjects 3 wk after initial sensitization generated secondary proliferative responses. To investigate the antigenic requirements for secondary stimulation, autologous TNP-conjugate primed responders were restimulated with both autologous and allogeneic TNP-conjugated stimulators. In all experiments restimulation with autologous conjugated cells yielded substantially greater proliferative responses than with allogeneic conjugates. Experiments were then performed to ascertain whether HLA determinant homology between primed responder and stimulator cells influenced the level of secondary responsiveness. Homology for HLA-A and B locus serologic determinants was not associated with enhanced responsiveness. In contrast, D region determinant homology, detected by B-cell antigen typing, showed a highly significant positive correlation with the magnitude of secondary responses. The data thus strongly suggest that for secondary proliferative responses to TNP, human T cells recognize hapten in association with HLA-D region determinants.